Loc device for amplifying and detecting target nucleic acid sequences using electrochemiluminescent resonant energy transfer, linear probes with covalently attached primers

Abstract

A lab-on-a-chip (LOC) device for amplifying and detecting target nucleic acid sequences, the LOC device having electrochemiluminescent (ECL), resonant energy transfer, linear probes for hybridization with the target nucleic acid sequences, each of the probes having a linear portion containing a sequence complementary to the target nucleic acid sequence, an ECL luminophore for emitting photons when in an excited state, a functional moiety for quenching photon emission from the ECL luminophore by resonant energy transfer, and a covalently attached primer for extension along a complementary sequence denatured from the target nucleic acid sequence to replicate the target nucleic acid sequence, heaters for thermally cycling the target nucleic acid sequences through a polymerase chain reaction (PCR), in which the covalently attached primers anneal to oligonucleotides containing the target nucleic acid sequences, and, electrodes for receiving an electrical pulse to excite the ECL luminophores, wherein during use, replicating the target nucleic acid sequence causes the linear portion to dissociate from the functional moiety such that the complementary nucleic acid sequence therein hybridizes to the target nucleic acid sequence and photons emitted by the ECL luminophore are not quenched.

Description

FIELD OF THE INVENTION[0001]The present invention relates to diagnostic devices that use microsystems technologies (MST). In particular, the invention relates to microfluidic and biochemical processing and analysis for molecular diagnostics.CO-PENDING APPLICATIONS[0002]The following applications have been filed by the Applicant which relate to the present application:GBS001USGBS002USGBS003USGBS005USGBS006USGSR001USGSR002USGAS001USGAS002USGAS003USGAS004USGAS006USGAS007USGAS008USGAS009USGAS010USGAS012USGAS013USGAS014USGAS015USGAS016USGAS017USGAS018USGAS019USGAS020USGAS021USGAS022USGAS023USGAS024USGAS025USGAS026USGAS027USGAS028USGAS030USGAS031USGAS032USGAS033USGAS034USGAS035USGAS036USGAS037USGAS038USGAS039USGAS040USGAS041USGAS042USGAS043USGAS044USGAS045USGAS046USGAS047USGAS048USGAS049USGAS050USGAS054USGAS055USGAS056USGAS057USGAS058USGAS059USGAS060USGAS061USGAS062USGAS063USGAS065USGAS066USGAS067USGAS068USGAS069USGAS070USGAS080USGAS081USGAS082USGAS083USGAS084USGAS085USGAS086USGAS087USGAS...

AI Technical Summary

Benefits of technology

[0095]The easily usable, mass-producible, inexpensive, compact, and light LOC device accepts a biological sample, amplifies the nucleic acid targets in the sample, identifies the sample's nucleic acid sequences via hybridization with electrochemiluminescence resonance energy transfer primer-linked linear probes using its integral image sensor, and provides the results electronically at its output pads, with the primer-linked linear probes providing for a large number of optimal parallel amplification reactions to b...

Problems solved by technology

Insufficient stringency can result in a high degree of nonspecific binding.

Excessive stringency can lead to a failure of appropriate binding, which results in diminished sensitivity.

Despite the advantages that molecular diagnostic tests offer, the growth of this type of testing in the clinical laboratory has been slower than expected and remains a minor part of the practice of laboratory medicine.

This is primarily due to the complexity and costs associated with nucleic acid testing compared with tests ba...

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